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Technical Bulletin
DNA IQ SystemSmall
Sample Casework Protocol
INSTRUCTIONS FOR USE OF PRODUCTS DC6700 AND DC6701.
PRINTED IN USA.
Revised 5/09
Part# TB296
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DNA IQ SystemSmall
Sample Casework Protocol
All technical literature is available on the Internet at: www.promega.com/tbs
Please visit the web site to verify that you are using the most current version of this
Technical Bulletin. Please contact Promega Technical Services if you have questions on use
of this system. E-mail: genetic@promega.com.
1. Description..........................................................................................................1
2. Product Components and Storage Conditions ............................................2
3. Sample Types Examined ..................................................................................5
4. Protocol for the DNA IQ System................................................................6
A.
B.
C.
D.
5. Troubleshooting...............................................................................................12
6. References .........................................................................................................13
7. Composition of Buffers and Solutions .......................................................13
8. Related Products ..............................................................................................14
1.
Description
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with silica matrices uses the affinity of DNA for silica and does not require
organic components. Silica filters are convenient when used with a filtration
system but tend to give lower yields and require extensive washing to remove
the guanidine-based lysis buffer. Currently available silica magnetic particles
tend to give higher yield but also need extensive washing.
The DNA IQ System(a) uses a novel paramagnetic resin for DNA isolation.
Using the DNA IQ System to process small casework samples requires two
steps. For biological material on solid supports, the first step provides an easy,
rapid, efficient and almost universal stain extraction method. This step is
unnecessary for liquid samples. The second step uses the paramagnetic resin to
purify DNA without requiring extensive washing to remove the lysis reagent.
This system is designed to rapidly purify small quantities of DNA and give
consistent yields for a specific sample type.
The DNA IQ System has been automated using the Beckman Coulter Biomek
2000 and 3000 Laboratory Automation Workstations and Tecan Freedom EVO
100 liquid handler. For more information, see the DNA IQ System product
profile at: www.promega.com/applications/hmnid/productprofiles/automation/.
For more information about implementing these methods, contact Promega
Technical Services (genetic@promega.com).
2.
Product
DNA IQ System
For Laboratory Use. This system includes:
3ml
150ml
70ml
50ml
0.9ml
40ml
30ml
15ml
Cat.#
DC6700
Size
100 samples
Cat.#
DC6701
Resin
Lysis Buffer
2X Wash Buffer
Elution Buffer
Product
DNA IQ System
For Laboratory Use. This system includes:
Size
400 samples
Resin
Lysis Buffer
2X Wash Buffer
Elution Buffer
Printed in USA.
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70C
65C
3237MB
Figure 1. Schematic of DNA isolation from stains on solid material using the
DNA IQ System. See Section 4.B for a detailed protocol.
Promega Corporation 2800 Woods Hollow Road Madison, WI 53711-5399 USA
Toll Free in USA 800-356-9526 Phone 608-274-4330 Fax 608-277-2516 www.promega.com
Printed in USA.
Revised 5/09
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65C
3238MB
Figure 2. Schematic of DNA purification from liquid samples using the DNA IQ
System. See Section 4.C for a detailed protocol.
Promega Corporation 2800 Woods Hollow Road Madison, WI 53711-5399 USA
Toll Free in USA 800-356-9526 Phone 608-274-4330 Fax 608-277-2516 www.promega.com
Part# TB296
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Printed in USA.
Revised 5/09
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DNA from the following sample types have been successfully purified at
Promega or by external forensic laboratories. Due to the nature of casework
samples (i.e, the samples may have been exposed to environmental factors for
long periods of time and the amount of biological material may be limiting),
DNA yields may vary, and DNA may not be obtained from all samples. Please
see the most updated list at: www.promega.com/dnaiqsamples/. Tissue
masses, including hair, bone, and sperm, require a proteinase K digestion to
obtain reliable amounts of DNA. Contact Promega Technical Services
(genetic@promega.com) for the latest information on available protocols.
Table 1. Types of Samples From Which DNA Has Been Successfully Isolated
Using the DNA IQ System.
Sample Type
Fresh blood
Frozen blood
Bloodstains
S&S 903 paper
FTA paper
Cotton
Blue denim
Black denim
Soil
Leather
Surface to swab
Buccal swabs
Cotton
Rayon
CEP paper
Swab to FTA paper
Foam swab to paper
Cigarette butt
Yes
Yes
Yes
Yes
Yes
Yes
Yes
Yes
Yes
Yes
Yes
Yes
Yes
Yes
Yes
Toothbrush
Yes
Envelope
Yes
Urine
Yes
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Yes
Bulletin TB307.
Formalin-fixed
Hair
Bone*
Antler*
Differential extractions
Yes
Yes
Yes
Yes
Yes
Yes
Yes
*Requires the Bone Incubation Buffer containing 1mg/ml proteinase K for DNA
purification. Please contact Technical Services (genetic@promega.com) for a protocol for
DNA isolation from bone samples.
4.
95100% ethanol
isopropyl alcohol
1M DTT
70C heat block, water bath or thermal cycler (for stain or swab extraction)
vortex mixer
Printed in USA.
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Total Volume
100l
100l
200l
1 Cotton swab
250l
100l
350l
250l
100l
350l
150l
100l
250l
150l
100l
250l
Cloth up to 25mm2
150l
100l
250l
1For
2For
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Printed in USA.
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7. Vortex tube for 2 seconds at high speed. Place tube in the magnetic stand.
Separation will occur instantly.
Note: If resin does not form a distinct pellet on the side of the tube, vortex
the tube and quickly place back in the stand.
8. Carefully remove and discard all of the solution without disturbing the
resin pellet on the side of the tube.
Note: If some resin is drawn up in tip, gently expel resin back into tube to
allow re-separation.
9. Add 100l of prepared Lysis Buffer. Remove the tube from the magnetic
stand, and vortex for 2 seconds at high speed.
10. Return tube to the magnetic stand, and discard all Lysis Buffer.
11. Add 100l of prepared 1X Wash Buffer. Remove tube from the magnetic
stand, and vortex for 2 seconds at high speed.
12. Return tube to the magnetic stand, and discard all Wash Buffer.
13. Repeat Steps 11 and 12 two more times for a total of three washes. Be sure
that all of the solution has been removed after the last wash.
14. With the tube in the magnetic stand and the lid open, air-dry the resin for
5 minutes.
Do not dry for more than 20 minutes, as this may inhibit removal of DNA.
15. Add 25100l of Elution Buffer, depending on how much biological material
was used. A lower elution volume ensures a higher final concentration of
DNA.
16. Close the lid, and vortex the tube for 2 seconds at high speed. Incubate the
tube at 65C for 5 minutes.
17. Remove the tube from the heat source, and vortex for 2 seconds at high
speed. Immediately place the tube on the magnetic stand.
Tubes must remain hot until placed in the magnetic stand or yield will
decrease.
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7l =
l of resin
(Number of samples + 1)
93l =
2. Mix liquid sample gently, and place an aliquot of up to 40l into a 1.5ml
microcentrifuge tube.
3. Vortex the resin/Lysis Buffer mixture for 3 seconds at high speed to ensure
suspension of resin, and add 100l of the mixture to the tube containing the
liquid sample. The resin/Lysis Buffer mixture should be mixed again if the
resin begins to settle while dispensing aliquots.
4. Vortex the sample/Lysis Buffer/resin mix for 3 seconds at high speed.
Incubate 5 minutes at room temperature. Vortex mixture for 3 seconds once
every minute during this 5-minute incubation.
5. Vortex for 2 seconds at high speed. Place tube in the magnetic stand.
Separation will occur instantly.
Note: If resin does not form a distinct pellet on the side of the tube, vortex
the tube and quickly place it back in the stand.
6. Carefully remove and discard all of the solution without disturbing the
resin pellet on the side of the tube.
7. Add 100l of prepared Lysis Buffer. Remove tube from the magnetic stand,
and vortex for 2 seconds at high speed.
8. Return tube to the magnetic stand, and remove and discard all Lysis Buffer.
9. Add 100l of prepared 1X Wash Buffer. Remove tube from the magnetic
stand, and vortex for 2 seconds at high speed.
10. Return tube to the magnetic stand. Dispose of all Wash Buffer.
11. Repeat Steps 9 and 10 two more times for a total of three washes. Be sure
all of the solution has been removed after the last wash.
12. With the tube in the magnetic stand and the lid open, air-dry the resin for
5 minutes.
Do not dry for more than 20 minutes, as this may inhibit removal of DNA.
Printed in USA.
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13. Add 25100l of Elution Buffer, depending on how much biological material
was used. A lower elution volume ensures a higher final concentration of
DNA.
14. Close the lid, and vortex tube for 2 seconds at high speed. Incubate at 65C
for 5 minutes.
15. Remove the tube from the heat source, and vortex for 2 seconds at high
speed. Immediately place on the magnetic stand.
Tubes must remain hot until placed in the magnetic stand, or yield will
decrease.
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5. Vortex for 3 seconds at high speed, and incubate at room temperature for
5 minutes.
6. Proceed to Steps 516 of Section 4.C to purify DNA from this nonsperm
fraction.
5.
Troubleshooting
For questions not addressed here, please contact your local Promega Branch Office or
distributor. Contact information available at: www.promega.com. E-mail: genetic@promega.com
Symptoms
Poor yield
Printed in USA.
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Troubleshooting (continued)
Symptoms
6.
References
1. Greenspoon, S. and Ban, J. (2002) Robotic extraction of mock sexual assault samples
using the Biomek 2000 and the DNA IQ System. Profiles in DNA 5, 35.
2. Gill, P. et al. (1985) Forensic application of DNA 'fingerprints'. Nature 318, 5779.
7.
10mM
100mM
50mM
0.5%
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Related Products
Product
MagneSphere Technology Magnetic
Separation Stand (two-position)
MagneSphere Technology Magnetic
Separation Stand (twelve-position)
PolyATtract System 1000 Magnetic
Separation Stand
DNA IQ Spin Baskets*
Microtubes, 1.5ml
ART 20P, Pipet Tip, 20l
ART 200, Pipet Tip, 200l
ART 1000E, Pipet Tip, 1,000l
Slicprep 96 Device**
AluQuant Human DNA Quantitation System*
Size
Cat.#
1.5ml
Z5332
1.5ml
Z5342
1 each
1,000/pk
1,000/pk
960/pk
960/pk
800/pk
10 pack
80 determinations
400 determinations
Tissue and Hair Extraction Kit (for use with DNA IQ)**
100 reactions
DTT, Molecular Grade (Dry Powder)**
5g
25g
PowerPlex 16 System*
100 reactions
400 reactions
PowerPlex 1.1 and 2.1 Systems*
100 reactions
400 reactions
PowerPlex 1.2 System*
100 reactions
Z5410
V1221
V1231
DY1071
DY1121
DY1131
V1391
DC1010
DC1011
DC6740
V3151
V3155
DC6531
DC6530
DC6501
DC6500
DC6101
(a)U.S. Pat. Nos. 6,027,945, 6,368,800 and 6,673,631, Australian Pat. No. 732756, European Pat. Nos. 0 895 546 and
1 204 741 and Mexican Pat. No. 209436 have been issued to Promega Corporation for methods of isolating biological target
materials using silica magnetic particles and simultaneous isolation and quantitation of DNA. Other patents are pending.
Printed in USA.
Revised 4/06